CultureofsingleCells(Bergmann’scellplatingtechnique)

In this technique, free cell are suspended in a liquid medium (if cell aggregates are there, the culture is
filtered), and a culture medium with agar (0.6-1%) is cooled and maintained at 35ºC in a water bath. Equal
volumes of these liquid and agar media are mixed and rapidly spread in a Petri dish, so that cells are evenly
distributed in a thin layer, after solidification. The Petri dishes are sealed with parafilm and examined with
inverted microscope to mark single cells (marking is done on outer surface of the dish). The plates are
incubated in dark at 25ºC and cell colonies developing from marked single class, are used to obtain
methods (e.g. filter paper raft-nurse technique; microchamber technique, etc.) have also been developed
to grow individual cells.

Bergamanns cell Plating Technique for culturing single cells

1. Fine gauze 2. Filtrate with single cells and cell 3. Molten White agar
aggregates medium
4. Cell suspension 5. Cell colony 6. Petri plate
7. Agar medium with single cells 8. Top view 9. Side view
Culture of Single Cells (Bergmanns Cell Plating Techniques) - In this technique, free cells are
suspended in a liquid medium (if cell aggregates are there, it is filtered). Culture medium with agar (0.6 -
1%) is cooled and maintained at 35°C in a water bath.

Equal volumes of liquid and agar media are mixed and rapidly spread in a Petri dish, so that cells are
evenly distributed in a thin layer, after solidification. The Petri dishes are sealed with parafilm and
examined with inverted microscope to mark single cells (marking is done on outer surface of the dish).

The Plates are incubated in dark at 25°C and cell colonies developing from marked single cells, are used to
obtain single cell cultures. Various other methods (e.g. filter paper raft nurse technique; microchamber
technique) have also been developed to grow individual cells (see Bhojwani and Razdan, 1983).